Genetically optimised microorganism for producing molecules of interest

Abstract

The invention concerns a genetically modified microorganism expressing a functional type I or II RuBisCO enzyme and a functional phosphoribulokinase (PRK), and in which the glycolysis pathway is at least partially inhibited, said microorganism being genetically modified so as to produce an exogenous molecule and/or to overproduce an endogenous molecule. According to the invention, the oxidative branch of the pentose phosphate pathway may also be at least partially inhibited. The invention also concerns the use of such a genetically modified microorganism for the production or overproduction of a molecule of interest and processes for the synthesis or bioconversion of molecules of interest.

The invention claimed is: 
     
       1. Genetically modified microorganism expressing a functional RuBisCO enzyme and a functional phosphoribulokinase (PRK), and in which glycolysis pathway is inhibited by inhibiting the expression of a gene encoding glyceraldehyde 3-phosphate dehydrogenase and/or a gene encoding phosphoglycerate kinase, and oxidative branch of pentose phosphate pathway of said microorganism is also inhibited by inhibiting the expression of a gene encoding glucose-6-phosphate dehydrogenase or 6-phosphogluconolactonase or 6-phosphogluconate dehydrogenase, said microorganism being genetically modified so as to produce an exogenous molecule of interest and/or to overproduce an endogenous molecule of interest, other than the RuBisCO or phosphoribulokinase enzyme. 
     
     
       2. The genetically modified microorganism of to  claim 1 , said microorganism being genetically modified to express a recombinant RuBisCO enzyme and/or PRK. 
     
     
       3. The genetically modified microorganism of to  claim 1 , said microorganism being genetically modified to inhibit the oxidative branch of the pentose phosphate pathway upstream of ribulose-5-phosphate production. 
     
     
       4. The genetically modified microorganism of to  claim 1 , wherein the exogenous molecule and/or the endogenous molecule is selected from the group consisting of amino acids, peptides, proteins, vitamins, sterols, flavonoids, terpenes, terpenoids, fatty acids, polyols and organic acids. 
     
     
       5. The genetically modified microorganism of  claim 1 , said microorganism being a eukaryotic cell selected from yeasts, fungi, microalgae, and a prokaryotic cell. 
     
     
       6. The genetically modified microorganism of  claim 1 , said microorganism being a yeast of the genus  Saccharomyces cerevisiae  genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the TDH1, TDH2 and/or TDH3 gene is inhibited. 
     
     
       7. The genetically modified microorganism of  claim 1 , said microorganism being a  Saccharomyces cerevisiae  yeast genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the PGK1 gene is inhibited. 
     
     
       8. The genetically modified microorganism of  claim 6 , wherein the expression of the ZWF1 gene is inhibited. 
     
     
       9. The genetically modified microorganism of  claim 1 , said microorganism being a filamentous fungus of the genus  Aspergillus  genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the pgk and gsdA genes is inhibited. 
     
     
       10. The genetically modified microorganism of  claim 1 , said microorganism being an  E. coli  bacterium genetically modified to express a functional type I or II RuBisCO and a functional phosphoribulokinase (PRK), and in which the expression of the gapA and/or pgk gene is inhibited. 
     
     
       11. A process for producing an amino acid, peptide, protein, vitamin, sterol, flavonoid, terpene, terpenoid, fatty acid, polyol or organic acid comprising (i) inserting at least one sequence encoding an enzyme involved in the synthesis or bioconversion of said amino acid, peptide, protein, vitamin, sterol, flavonoid, terpene, terpenoid, fatty acid, polyol or organic acid into the genetically modified microorganism of  claim 1 , (ii) culturing said microorganism under conditions allowing expression of said enzyme and optionally (iii) recovering and/or purifying said amino acid, peptide, protein, vitamin, sterol, flavonoid, terpene, terpenoid, fatty acid, polyol or organic acid.